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Figure 4 | Molecular Cancer

Figure 4

From: The monoclonal antibody Zt/f2 targeting RON receptor tyrosine kinase as potential therapeutics against tumor growth-mediated by colon cancer cells

Figure 4

Effect of Zt/f2 on RON expression by colon cancer cells. A) Down-regulation of RON/RON160 expression by Zt/f2. HT-29 cells (1 × 106 cells/sample) were treated with Zt/f2 (10 μg/ml) or control mouse IgG at 37°C for 6 and 12 h. After treatment, cells were washed with acidic buffer to eliminate surface-bound antibody (Guin et al., 2010) and then incubated with Zt/g4 (1 μg/sample) to detect RON/RON160 remaining on the cell surface by flow cytometric analysis. B) Preventive effect of filipin on Zt/f2-induced RON reduction. HT-29 cells were pre-incubated with filipin (10 μg/ml) for 30 min and then treated at 37°C for 12 h with Zt/f2 or control mouse IgG (10 μg/ml). After washing with acidic buffer, RON on cell surface was detected by Zt/g4 as previously described (Guin et al., 2010). C) Kinetic effect of Zt/f2 on RON/RON160 expression. HT-29 cells were treated with 10 μg/ml of Zt/f2 for various times. RON expression was determined by Western blot analysis. Normal mouse IgG was used as the control. D) Preventive effect of lactacystin on Zt/f2-induced RON reduction. HT-29 cells were pre-incubated with 1 or 10 μg/ml of lactacystin for 30 min and then treated with 10 μg/ml of Zt/f2 at 37°C for 12 h. Levels of RON were determined by Western blot analysis.

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